Nextgen Dissection of the Genomic Basis of Kidney Development

描述（由申请人提供）：本提案的长期目标是对驱动肾脏发育的基因表达程序进行基于NexGen RNA-Seq的解剖。中心论点是直截了当的。我们建议定义，在一个敏感的和定量的方式，所有的基因组活跃在每个关键细胞类型的发育肾脏。将生成一个图谱，全面的特征，包括microrna和更正统的编码mrna，并提供详细的分析替代RNA加工事件。最终的结果将是一个重要的资源，描述模拟基因表达代码，定义和驱动发育中的肾脏的每个隔室。这个资源将确定所有的生长因子，受体和转录因子表达在每种细胞类型。它将建立一套新的细胞类型特异性分子标记，用于突变分析。此外，它将为全球分析由于突变或疾病而发生的基因表达模式变化提供一个标准基线。此外，我们建议使用Chip-Seq方法来发现转录因子Hoxa11， Lhx1和Six2的直接下游靶点。每一种都被证明在肾脏发育中发挥重要作用，通过确定它们的直接下游靶点，我们将更好地了解它们在肾脏形成过程中的确切作用。

DESCRIPTION (provided by applicant): The long-term objective of this proposal is to conduct a NexGen RNA-Seq based dissection of the gene expression program driving kidney development. The central thesis is straightforward. We propose to define, in a sensitive and quantitative manner, all of the sets of genes active in each critical cell type of the developing kidney. An atlas will be generated, comprehensive in character, including microRNAs as well as more orthodox coding mRNAs, and providing detailed analyses of alternative RNA processing events. The end result will be an essential resource, describing the analog gene expression code that defines and drives each compartment of the developing kidney. This resource will identify all of the growth factors, receptors and transcription factors expressed in each cell type. It will establish a set of novel cell type specific molecular markers, useful in mutant analyses. In addition it will provide a standard baseline for the global analysis of changes in gene expression patterns occurring as a result of mutation or disease. In addition we propose to use a Chip-Seq approach to discover the direct downstream targets of the transcription factors Hoxa11, Lhx1 and Six2. Each has been shown to play an important role in kidney development, and by identifying their direct downstream targets we will gain a better view of their precise roles in the process of nephrogenesis. PUBLIC HEALTH RELEVANCE: The proposed work would use the latest revolutionary sequencing technology to characterize all of the gene activities in the many different cell types of the developing kidney. This would lead to a much deeper understanding of the genetic program that drives the process of making a kidney, which in turn will in time help us better understand how to repair kidneys that have been damaged by disease.